KR20080110852A - Endothelin and Endothelin Receptor Agonists for the Treatment of Metabolic Disease - Google Patents

Abstract

Methods for treating a condition or disease that can be alleviated by reducing food intake include administering an effective amount of an endothelin or endothelin agent, alone or in combination with other compounds or compositions that may affect satiety. The methods of the invention are useful for treating conditions or diseases including obesity, type II diabetes, eating disorders, and insulin-resistant syndrome. Pharmaceutical compositions useful for use in the methods of the invention are described herein.

Description

ENDOTHELIN AND ENDOTHELIN RECEPTOR AGONISTS IN THE TREATMENT OF METABOLIC DISEASES}

References related to this application

This patent claims priority to US Patent Application No. 60 / 785,447, filed March 23, 2006, which is incorporated herein by reference in its entirety.

The present invention relates to the field of medicine and health. More specifically, the present invention relates to methods and compositions that utilize at least one endoterin or endothelin receptor agonist to reduce food intake, lose weight, or treat metabolic diseases such as diabetes and obesity.

Obesity affects millions of people, especially in developed countries like the United States. According to the World Health Organization, more than one billion adults worldwide are overweight, at least 300 million of whom are obese (body mass index (BMI) above 30). See Smith et al., Nature Medicine 12: 75-80 (2005). In the United States alone, about 65% of adults are overweight or obese. Surprisingly, overweight and obesity account for about 10% of children worldwide. See Yach et al., Nature Medicine 12: 62-66 (2005). Being overweight has psychological consequences such as cardiovascular disease, stroke, osteoarthritis, type 2 diabetes, sleep apnea, respiratory disease, gastroesophageal reflux disease, metabolic syndrome, cancer as well as low self-esteem and clinical depression Including a myriad of conditions and diseases are susceptible. Also important is that obesity can actually significantly increase morbidity and mortality from all other diseases.

Although the onset of obesity is often due to a variety of reasons, the ingestion of excessively high calories (ie food or nutrients) relative to energy consumption is the main cause. Excess calories cause increased fat or lipid tissue production. Maintaining this reduction of adipose tissue can reduce or even increase the devastating consequences of obesity. In general, limiting total calorie intake or increasing energy consumption, such as increasing exercise, can reduce adipose tissue.

Despite the simplicity of solving the problem, it is extremely difficult to steadily reduce the weight of overweight and obese patients, especially in obese patients. Therapeutic methods include diet and exercise therapy, behavioral change techniques and medications such as appetite suppressants and food absorption inhibitors, from gastric bypass surgery and physical equipment such as jaw wire fixation, waist cords, and balloons. It varies. After all, the purpose of these methods is to steadily reduce food or nutrient absorption. However, various factors make it difficult to achieve the goal of consistently reducing food intake. Some treatments are not suitable for high obesity. For example, these patients are subjects who are too large to exercise safely or which are not suitable for surgical procedures. In addition, a decrease in body fat is necessary, but a decrease in lean body weight is not necessary. The lean body weight consists of muscles, living organs, bones, muscle tissue and other non-fat tissues, the reduction of which is detrimental to the health of the individual. Still, many therapies are weakened by these therapies because they result in a decrease in fat tissue and lean body mass at the same time. Additional factors make it difficult to continuously reduce food intake, including significant calorie restriction followed by increased appetite and reduced energy levels.

Endothelin is an important factor in maintaining health and the cardiovascular system. See, eg, Miyauchi, Ann. Rev. Physiol . 61: 391-415 (1999). One group uses methods and compositions to block or counteract endothelin expression by interfering with the enzymatic activity of endothelin-converting enzyme-2 (ECE-2), such as obesity and diabetes. It has been proposed to target the endothelin system for the treatment of metabolic diseases. See, eg, US Patent Application No. 2003/0232044. In other words, treatment of metabolic disease is realized by reducing or increasing endothelin activity using modulators of ECE-2 activity.

As obesity continues to increase worldwide, there is a great need for a much more effective way to reduce food intake and lead to weight loss as well as to maintain weight loss for extended periods of time. Endothelin and endothelin receptor agonists provide new therapeutic alternatives to conditions and diseases that can benefit from reduced food intake.

Summary of the Invention

Provided herein are foods, including methods of administering an effective amount of an endoterin or endothelin agonist alone or in combination with other compounds and compositions that reduce food intake, increase satiety or increase or maintain weight loss. Methods and compositions for treating conditions or diseases that can be alleviated by reducing intake. This method is effective for treating conditions including obesity, type II diabetes, metabolic syndrome and insulin-resistant syndrome.

Accordingly, provided in one aspect herein is a method of reducing food intake in a patient who desires or needs to reduce food intake by administering an effective dose of an endoterin or endothelin agonist to reduce food intake.

Provided in another aspect is a method of reducing or maintaining the weight of a patient in need or need to lose or maintain weight, comprising administering an effective dose of an endoterin or endothelin agonist to reduce or maintain weight .

Further provided herein is a method of treating obesity in a patient requiring or requiring treatment for obesity, comprising administering an effective dose of an endoterin or an endothelin agonist to treat obesity.

Also provided herein is a patient in need or need for the prevention or treatment of a metabolic disease, comprising administering an effective amount of an endoterin or endothelin agonist to reduce or alleviate at least one symptom of the metabolic disease. To prevent or treat metabolic diseases. In one embodiment, the metabolic disease is obesity, diabetes, insulin-dependent syndrome, syndrome-X, or a disease associated with excessive calorie intake.

In one embodiment of the method, the endothelin is for example endotherin-1 of SEQ ID NOs: 1-6; Eg, endoterin-2 of SEQ ID NOs: 7-9; Or at least one of endorin-3 of SEQ ID NOs: 10-13. In one embodiment, the endoterin is endoterin-3. In other embodiments, the endoterin is any one of SEQ ID NOs: 1, 7, 10 or a combination thereof. In other embodiments, the endothelin is any one of SEQ ID NOs: 1-13 or a combination thereof, including each subset specifically excludes one or more of SEQ ID NOs: 1-13. In another embodiment, the endoterin has an amino acid sequence having sequence homology of at least 75%, at least 80%, at least 85%, at least 90% or 95% with any one of SEQ ID NOs: 1, 7 or 10. In other embodiments, the endothelin agonist is an endothelin agonist that does not comprise at least 5 or at least 10 amino acid substitutions, deletions, or additions as compared to any one of SEQ ID NOs: 1, 7 or 10. In other embodiments, the endotherin agonist is an endotherin agonist that does not comprise at least 5 amino acid substitutions, deletions, or additions as compared to any one of SEQ ID NOs: 1, 7 or 10.

In another embodiment, the endothelin agonist is a fragment of endothelin-1, endothelin-2, or endothrin-3, wherein the fragment binds to and activates an ET _A or ET _B receptor. In another embodiment, the fragment is preferably bound to and activated by the ET _B receptor; In one embodiment, the fragment binds to and activates the ET _B receptor, but does not activate the ET _A receptor. In one embodiment, the fragment does not comprise at least 7 amino acid deletions compared to, for example, the full length endoterin of SEQ ID NOs: 1-13.

In another embodiment, the endothelin receptor agonist is sarafotoxin, for example any one of SEQ ID NOs: 15, 16, 17, 18, 31 or 32. Also included are all combinations of SEQ ID NOs: 15, 16, 17, 18, 31 or 32 which include all subsets which may specifically exclude one or more of the sequences. In other embodiments, sarapotoxin has at least 80%, at least 85%, at least 90% or 95% amino acid homology with any one of SEQ ID NOs: 15, 16, 17, 18, 31 or 32. In other embodiments, sarapotoxin does not comprise at least 10 or at least 5 amino acid deletions, additions or substitutions as compared to any one of SEQ ID NOs: 15, 16, 17, 18, 31 or 32. In one embodiment, sarapotoxin preferably binds to and activates the ET _B receptor.

In another embodiment, the endothelin agonist is demaseptin, and in more specific embodiments the demaceptin is adenoregulin. In one embodiment, demasceptin or adenoregulin comprises SEQ ID NOs: 20, 21, 22, 23, 24, 25, 26, 27, comprising all subsets that may specifically exclude one or more of SEQ ID NOs: 20-30 28, 29, 30, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, Amino acid sequence of any one of 55, 56, 57, 58, 59, 60, 61, 62, 63, 64 or a combination thereof. In another embodiment, demasceptin or adenoregulin has an amino acid homology of at least 75%, at least 80%, at least 85%, at least 90% or 95% with any one of SEQ ID NOs: 20-30 or 33-64. In other embodiments, derma septin or adenoregulin does not comprise at least 10 or at least 5 amino acid deletions, additions or substitutions as compared to any one of SEQ ID NOs: 20-30 or 33-64. In one embodiment, demasceptin or adenoregulin preferably binds to and activates the ET _B receptor.

In addition, the methods disclosed herein may further comprise administering the compound, which compound increases satiety, reduces food intake or increases or maintains weight loss. Such compounds include, but are not limited to, exendin, amylin, PYY, leptin, oxyntomodullin, neuroromedin or cholecystokinin (CCK), Agonist analogs or derivatives thereof. In certain embodiments, the endothelin agonist can be adenoregulin, IRL1620, sarapotoxin, or any combination thereof.

In one embodiment, the endothelin or endothelin agonist acts through the ET _B receptor. In certain embodiments, the endothelin or endothelin agonist does not act through the ET _A receptor.

Further provided herein include at least one of an endoterin or an endoterin agonist for treating a condition or disease that may be alleviated by reducing food intake as provided herein in a dose effective to treat the condition. Use of the formulation. Also provided are uses for the manufacture of a drug for treating or affecting a disease or condition as disclosed herein, of at least one of an endoterin or an endothelin agonist.

1 shows a dose-dependent decrease in food intake 120 minutes after administration of the endothelin receptor B (ET _B ) agonist.

2 shows a dose-dependent decrease in food intake 30 minutes after administration of the ET _B agonist.

Figure 3 shows a dose-dependent decrease in food intake 60 minutes after administration of the ET _B agonist.

FIG. 4 shows body fat when adenoregulin is administered to diet-induced obesity (DIO) mice treated with medium only and maintained on high fat diet or mice fed low fat diet and treated only medium mediated. Shows a decrease.

5 shows weight loss in diet-induced obesity (DIO) mice treated with 300 nmol / kg / d of adenoregulin for 28 days and maintained on a high fat diet. HF saline: DIO mice maintained on a high fat diet and treated with mediators only (ie saline). LF Saline: DIO mice maintained on a low fat diet and treated with media only. DIO mice were maintained on a high fat diet serving as a positive control and treated with peptide YY (PYY).

6 shows weight loss in DIO mice treated with 20 nmol / kg / d endotherin 1 (ET-1) for 28 days and maintained on a high fat diet via a subcutaneous (s.c) osmotic pump.

Provided herein is the use of endoterin and endothelin receptor agonists for reducing food intake or for reducing food intake or for weight loss or maintenance of a patient who desires or needs weight loss or maintenance. Endoterin effectively provides activity in a number of normal physiological and pathophysiological conditions. See, eg, Kezierski et al., Ann. Rev. Pharmacol. Toxicol. 41: 851-76 (2001). Endoterin acts as an effective perisecreting vasoconstrictive peptide that maintains basal vascular tone and regulates blood vessel growth. These peptides regulate lung respiration and blood vessel tension, regulate kidney fluid and sodium secretion, as well as acid-base balance and neurotransmitters. Endothelin is also involved in the development and pathology of hypertension, arteriosclerosis, cardiac hypertrophy, congestive heart disorders, pulmonary arterial hypertension and kidney disorders. Factors that promote endothelin production and activity include thrombin, TGF-β, TNF-α, norepinephrine and insulin. See, eg, Miyauchi et al., Ann. Rev. Physiol . 61: 391-415 (1999). However, to date, the function of endoterin in obesity and diabetes is associated with the regulation of vascular complications and hypertension observed in patients with these diseases. See, eg, Wolfer et al., Metabolism 42: 1027-30 (1993); Ferri et al., Exp. Clin. Endocrinol. Diabetes 105 (S2): 38-40 (1997).

Provided herein is the discovery that systemic administration of endothelin acts to reduce food intake, lose weight, or both. Thus, methods for losing weight using endothelin and endothelin receptor agonists may include metabolic diseases such as obesity, type II diabetes, insulin resistance syndrome, syndrome X, metabolic syndrome or other diseases associated with undesirable or excessive calorie intake. Shows a new therapeutic approach to treat Reducing food intake in such patients may be useful in reducing plasma glucose levels, plasma lipid levels, and cardiovascular disease.

Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All patents, applications, publications, and other publications mentioned herein are incorporated by reference in their entirety. If the definitions described in this section are contrary to or inconsistent with the definitions described in patents, applications, publications, and other publications incorporated herein by reference, the definitions described in this section are equivalent to those incorporated herein by reference. It is superior to justice.

As used herein, "one" means "at least one" or "one or more".

The term "a condition or disease that can be treated, alleviated or suppressed by reducing food intake" can be alleviated or avoided by relatively high food intake, worsened or worsened, or reduced by food intake and / or weight loss. It refers to any condition or disease of a patient that can. Such conditions or diseases include, but are not necessarily limited to, obesity, diabetes, including type II diabetes, insulin-resistant syndrome, and syndrome X. Reducing food intake can help you to control, reduce or maintain your weight and work with your diet plan to reduce your daily calorie intake.

The term “endotherin agonist” refers to any isolated, naturally occurring or synthesized compound that binds to the endothelin receptor and results in a reduction in food intake by mimicking the reduced food intake of endothelin at the endothelin receptor. It is called. Thus, for the purposes of the present invention, endothelin agonists and endothelin receptor agonists are used interchangeably. In one embodiment, the endothelin agonist will specifically mimic the action of one or more of endothelin-1, endothelin-2, or endothelin-3. Biological actions include initiating one or more signaling components, including, for example, PLC activation, phospholipase A ₂ activation, phospholipase D activation, MAPK activation, pro-tumor gene expression (eg , c- fos, c-myc, c-jun ) or Shc-Grb2 complex formation. Such activity can be confirmed in vitro or in vivo by standard methods and assays. See, eg, Badr et al., J. Clin. Invest . 83: 336-42 (1989); Takuwa et al., J. Clin. Invest . 85: 653-58 (1990); Resnik et al., Eur. J. Biochem . 189: 415-21 (1990); Cazaubon et al., J. Biol. Chem . 269: 24805-09 (1994); Wang et al., Am. J. Physiol . 267: C1130-35 (1994); Simonson et al., J. Biol. Chem . 267: 8643-49 (1992).

As used herein, the term “obesity” or “overweight” refers to a person having a body mass index (BMI) of 30 kg / m ² or greater. See, eg, National Institute of Health, Clinical Guidelines on the Identification, Evaluation, and Treatment of Overweight and Obesity in Adults (1998). The term “overweight” refers to a person whose body mass index (BMI) is 25 kg / m ² or more, but who wants to lose weight regardless of his or her BMI or less than 30 kg / m ² . BMI is a value representing the weight-to-height relationship (or ratio) based on the weight (kg) of an individual divided by the square of its height (meters) (ie wt / (ht) ² ).

The term “metabolic disease” refers to diseases, conditions and conditions caused or characterized by abnormal energy use or consumption in the body. Examples of metabolic diseases include, but are not limited to, obesity, type II diabetes, insulin resistance syndrome, metabolic syndrome and syndrome X.

As used herein, the term “subject” refers not only to humans, but also to livestock and farm animals, non-human primates, and zoo animals, sport animals or pets and other mammals such as dogs, cats, horses, cows, and the like. Include. Those skilled in the art will readily appreciate that such a patient is a person who requires or needs a reduction in food intake, weight loss, or maintenance of reduced weight. In certain embodiments, the patient is a human.

The term “treating” or “treatment” is intended to alleviate, alleviate, alleviate, change, treat, ameliorate, enhance, influence, slow or stop the progression of a disease, at least one condition or Refers to the administration of one or more therapeutic agents to a patient who has a condition or disease that is at the manifestation of a disease or is more likely to develop a condition or disease, or who is more likely to develop a condition or disease.

The endothelin system comprises three endothelins, two endothelin receptors and two endothelin specific proteases. Endoterin is present in one of three isoforms: endoterin-1 (ET-1), endoterin-2 (ET-2), or endoterin-3 (ET-3). Each endothelin starts with a preproendothelin consisting of about 200 amino acid residues. Furin-like endopeptidase cleaves the bibasic position of the endothelin precursor to form a biologically inactive intermediate of about 37-41 amino acids, known as big ETs or pro-endotherins. do. And endoterin-converting enzymes (eg, ECE-1 and ECE-2) cleave big ETs to form a biologically active final product of 21 amino acids. Mammals express two endothelin receptors, ET _A and ET _B. These G-linked receptors have seven transmembrane domains between about 400 amino acid sequences thereof and undergo a signaling process involving phospholipase C-β activation, an increase in intracellular Ca ²⁺ and immediate initial gene expression. Induce. These receptors show different affinities for the three ETs. ET _A has nanomolar levels of affinity (20-60 pm) for ET-1 and ET-2, but not for ET-3 (6500 pm). ET _B shows equivalent affinity for ET-1, ET-2 and ET-3. See, eg, Arai et al., Nature 348: 730-32 (1990); Sakurai et al., Nature 348: 732-35 (1990), Sakurai et al., TiPS 13: 103-08 (1992); See US Pat. No. 6,821,743.

The ET-3 / ET _B system seems to be functionally different from the ET-1 / ET _A system. The ET-1 / ET _A system appears to induce a proliferative and vasoconstrictive effect on ET _A expression observed through vasculature. Thus, ET _A antagonists are often used to repair hemostasis. In contrast, ET _B expression appears to be restricted only in the stomach, pituitary gland, brain gland, submandibular gland, brain, heart, kidney, tachycardia and stomach. For example, Matsmoto et al., Biochem. Biophys. Res. Commun. 164: 74-80 (1989); Shiba et al., Biochem. Biophys. Res. Commun 186: 588-94 (1992). ET-3 / ET _B primarily induces vasodilation but in some situations also appears to induce vasoconstriction and cell proliferation.

In general, the level of circulation of endothelin is lower than that required for biological activity. For example, local ET-1 concentration in the vessel wall is ≧ 100 times the plasma level. See, eg, Kezierski et al., Ann. Rev. Pharmacol. Toxicol. 41: 851-76 (2001). The apparent need for high local concentrations for the vascularizing activity of endothelin allows systemic administration of doses of endothelin antagonists and agents to show the benefits of other effects of endothelin. In one example, endothelin agonists succeed in increased chemotherapy-induced tumor suppression in animals without systemic cardiovascular side effects following systemic administration. See, eg, US Patent Application No. 2004/0138121; See Rajeshkumar et al., Breast Cancer Res . 94: 237-47 (2005).

All suitable sources of ET-1, ET-2 or ET-3 can be used in the ET of the process of the invention. ETs can be isolated or purified from naturally occurring sources and can be prepared using molecular biological methods (eg, exogenous preparation), or chemical synthesis. In one embodiment, ETs useful in the compositions and methods of the present invention do not include big ETs (pro-ETs) or ETs precursors. In other embodiments, ETs useful in the methods and compositions of the present invention have an amino acid sequence comprising 36 or fewer amino acids. Examples of endothelin useful in the methods and compositions of the present invention are listed in Table 1. In general, endoterins comprise four cysteine residues that form two sets of disulfide bonds between two outer and two inner cysteine pairs. For example, the disulfide bond in SEQ ID NO: 1 may be present between ¹ C and ¹⁵ C and between ³ C and ¹¹ C. In one embodiment, the endothelin and the endothelin agonist may be provided to the patient in an inactivated (prodrug-drug) form comprising the endothelin, sarapotoxin, demasceptin or adenoregulin as disclosed herein. Suitable pro-drug forms include endoterin precursors or big ETs.

Examples of ET _B agents include, but are not necessarily limited to, agents, derivatives or functional fragments of ET-1, ET-2 or ET-3. In addition, representative sequences of endoterin are not necessarily limited to those described in Inoue et al., Proc. Natl. Acad. Sci. USA 86: 2863-67 (1989); And US Pat. No. 5,294,569; 5,231,166; 5,231,166; And 4,981,950, which are incorporated by reference. These agents, derivatives or fragments may be prepared using standard molecular biology techniques or chemical synthesis, which maintain biological functions or activities such as reducing food intake, weight loss or both. In one embodiment, the agent binds to the ET _B receptor. In certain embodiments, the agent selectively binds to the ET _B receptor but does not bind to the ET _A receptor sufficiently to elicit detectable ET _A activity in standard assays. In one embodiment, the agent does not bind to the ET _A receptor. In one embodiment, the endothelin agonist binds to the receptor with much greater affinity than the receptor, eg, at least 5, 10, 25, 100, 200, or 1000 times greater affinity.

ET _B agonists useful in the methods of the invention are not necessarily limited to BQ-788 (Clinalfa AG; N-cis-2,6-dimethylpiperidinocarbonyl-L-γ-methylleucil-D-1-me Oxycarbonyltryptophanyl-D-norleucine; see Ishikawa et al., Proc. Natl. Acad. Sci. USA 91: 4892 (1994)), IRL1620 (N-succinyl- [Glu ⁹ , Ala] ^11,15 ] ^-Endoterin 1 fragment 8-21; see Takai, et al., Biochem. Biophys. Res. Commun. 184: 953 (1992)), BQ-3020 (N-acetyl- [Ala ^{11 , 15} ] -Endoterin 1 fragment 6-21; see Ihara, et al., Life Sci. 51: PL47-PL52, (1992)), and [Ala ^{1, 3, 11, 15} ] ET-1 (ASASSLMDKEAVYFAHLDIIW [SEQ ID NO: 14]).

Another source of ET _B agonists is sarapotoxin. Separate from the dormant species (burrowing asp) the acids den art rock Tasman pick yen (Atractaspic engadensisan), the bulb-coagulant toxin is preferably combined with ET _B. See Takasaki et al., Toxicon 26: 543-48 (1988). Examples of sarapotoxin ET _B agonists are listed in Table 2 below. Further sarapotoxins include CSCNDMNDKECMYFCHQDVIW (SEQ ID NO: 31) and CSCKDMSDKECLNFCHQDVIW (SEQ ID NO: 32). As with endoterin, disulfide bonds may be present between the outer and inner pairs of sarapotoxin cysteine residues.

Examples of Sarapotoxin ET _B Agonists Sarapotoxin S6a c (Cys-Ser-Cys-Lys-Asp-Met-Thr-Asp-Lys-Glu-Cys-Leu-Asn-Phe-Cys-His-Gln-Asp-Val-Ile-Trp) [SEQ ID NO: 15] Sarapotoxin S6b c (Cys-Ser-Cys-Lys-Asp-Met-Thr-Asp-Lys-Glu-Cys-Leu-Tyr-Phe-Cys-His-Gln-Asp-Val-Ile-Trp) [SEQ ID NO: 16] Sarapotoxin S6c c (Cys-Thr-Cys-Asn-Asp-Met-Thr-Asp-Glu-Glu-Cys-Leu-Asn-Phe-Cys-His-Gln-Asp-Val-Ile-Trp) [SEQ ID NO: 17] Sarapotoxin S6d c (Cys-Thr-Cys-Asn-Asp-Met-Thr-Asp-Lys-Glu-Cys-Leu-Tyr-Phe-Cys-His-Glu-Asp-Ile-Ile-Trp) [SEQ ID NO: 18]

In another embodiment, the ET receptor agonist is adenoregulin. Donly et al., Proc. Natl. Acad. Sci USA 89: 10960-963 (1992); Amy et al., Biochem. Biophys. Res. Comm. 191: 983-90 (1993). Known sequences of adenoregulin include gene bank sequences X70278 and X72387. Adenoregulin, isolated from the skin of the frog Phyllomedusa bicolor , belongs to the demasceptin family, which is an antibacterial peptide. See, eg, Amich et al., J. Biol. Chem . 269: 17847-52 (1994); Zhou et al., Biotechnol. Lett. 27: 725-30 (2005). Examples of adenoregins are shown in Table 3. An example of reducing food intake by further administering adenoregulin is shown in Table 4. In one embodiment, the endothelin receptor agonist is at least 15%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at 120 minutes post-dose, compared to the mediator or untreated group, Or at least 70% reduction.

Example of adenoregulin SEQ ID NO: 20 MAFLKKSLFLVLFLGLVSLSICEEEKRENEDEEEQEDDEQSEMKRGLWSKIKEVGKEAAKAAAKAAGKAALGAVSEAVGEQ SEQ ID NO: 21 MDILKKSLFLVLFLGLVSLSICEEEKRENEDEEKQDDEQSEMKRAMWKDVLKKIGTVALHAGKAALGAVADTISQGEQ SEQ ID NO: 22 MAFLKKSLFLVLFLGLVSLSICEEEKRENEDEEEQEDDEQSEMKRGLWSKIKEAGKAALTAAGKAALGAVSDAVGEQ SEQ ID NO: 23 MAFLKKSLFLVLFLGLVSLSICEEEKRENKDEIEQEDDEQSEEKRALWKDILKNVGKAAGKAVLNTVTDMVNQGEQ SEQ ID NO: 24 MASLKKSLFLVLFLGLVSLSICEEEKRENEDEEEQEDDEQSEMKRGLWSNIKTAGKEAAKAALKAAGKAALGAVTDAVGEQ SEQ ID NO: 25 MDILKKSLFLVLFLGLVSLSICEEEKRENEDEEKQDDEQSEMKRAMWKDVLKKIAGKAALGAVADTISQGEQ SEQ ID NO: 26 MAFLKKSLFLVLFLGLVSLSVCEEEKRENEDEMEQEDDEQSEEKRALWKDILKNAGKAALNEINQLVNQGEL SEQ ID NO: 27 MAFLKKSVFLVLFLGLVSLSICEEEKREEENEEKQEDDEQSEEKRALWKNMLKGIGKLAGQAALGAVKTLVGAE SEQ ID NO: 28 MAFLKKSLFLVLFLGLVPLSLCESEKREGENEEEQEDDQSEEKRSLGSFLKGVGTTLASVGKVVSDQFGKLLQAGQG SEQ ID NO: 29 GLWNKIKEAASKAAGKAALGFVNEMV SEQ ID NO: 30 DVLKKIGTVALHAGKAALGAVADTISQ

Additional ET receptor agonists can be readily identified using methods known in the art. See, eg , Davenport, Pharmacological Rev. 54: 219-26 (2002); See US Pat. No. 6,821,743. For example, an agent can be identified for endothelin-inducing activity in cells expressing ET _A , ET _B , or both using known in vitro or in vivo assays. Such cells can express receptors endogenously or exogenously using known molecular biology techniques. In one implementation, the ET _B selective agonist is not coupled to the ET _A does not induce its activity. On the other hand, in another implementation, the ET _A selective agonists are not bound to ET _B does not induce its activity.

Such agents may include a number of chemical classes. In certain embodiments, it is an organic molecule, preferably a small organic compound having a molecular weight of at least 50 and less than 2,500 Daltons. In addition, ET receptor agonists include biomolecule-like antibodies, peptides, polypeptides, peptide analogs, saccharides, fatty acids, steroids, purines, pyrimidines, analogs, structural agents, or combinations thereof. In one embodiment, the ET receptor agonist may comprise protein preparations or binding fragments or analogs thereof, such as peptides and antibodies, such as Fv, F (ab ') ₂ and Fab. Known pharmacokinetic agents can be direct or random chemical modifications such as acylation, alkylation, esterification, amidation, etc. to prepare structural agents that act as non-selective or selective ET receptor agonists.

In one embodiment, the ET receptor agonist is an endoterin variant such as an ET-1, ET-2, or agonist or functional fragment of ET-3. Functional fragments are endoterins that have less than 21 amino acids but which do not bind and activate the endothelin receptor and thus do not reduce food intake and / or weight. Analogues not only have five or fewer amino acid substitutions, but also at least 75%, 80%, 85%, 90%, or 95% amino acid sequence homology with the sequence of SEQ ID NO: 1, SEQ ID NO: 7 or SEQ ID NO: 10 It includes having. In other embodiments, the endothelin variant has at least 10, at least 7, at least 5, at least 4, amino acid deletions, additions or substitutions as compared to known endoterins such as SEQ ID NO: 1, SEQ ID NO: 7 or SEQ ID NO: 10 At least 3, at least 2 or at least one amino acid deletion, addition or substitution. Also contemplated are the ET-1, ET-2, and ET-3 sequences from species other than humans.

In one embodiment, the ET receptor agonist is a variant such as an agonist or functional fragment of sarapotoxin or adenoregulin in Table 2 or 3 herein. Functional fragments may comprise up to 21 amino acids, but may bind to and activate endothelin receptors to reduce food intake and / or weight. Analogs comprising them not only have up to 5 amino acid substitutions, but also at least 75%, 80%, 85%, 90%, or 95% sequence homology with the sequences in Table 2 or Table 3. In another embodiment, the variant does not comprise at least 10, at least 7, at least 5, at least 4, at least 3, at least 2 amino acid deletions, additions or substitutions as compared to the sequences in Table 2 or Table 3 Only single amino acid deletions, additions or substitutions. Also contemplated are variant sequences from species other than humans.

As is well known in the art, “sequence homology” is a relationship between two or more polypeptide sequences or two or more polynucleotide sequences, determined by comparing sequences. In the art, “homology” refers to the degree of sequence relationship between polypeptide or polynucleotide sequences, as determined by the match between sequence strands. Homology is not necessarily limited to this, but is described in Computational Molecular Biology , Lesk, AM, ed., Oxford University Press, New York (1988); Biocomputing: Informatics and Genome Projects , Smith, DW, ed., Academic Press, New York, 1993; Computer Analysis of Sequence Data, Part I , Griffin, AM and Griffin, HG, eds., Humana Press, New Jersey (1994); Sequence Analysis in Molecular Biology , von Heinje, G., Academic Press (1987); Sequence Analysis Primer , Gribskov, M. and Devereux, J., eds., Stockton Press, New York (1991); And Carillo, H., and Lipman, D., SIAM J Applied Math , 48: 1073 (1988). The method for confirming homology is designed to give the largest match between the sequences tested. In addition, methods for confirming homology are summarized in the commonly available programs. Computer programs that can be used to confirm homology between two sequences include, but are not limited to: GCG (Devereux, J., et al., Nucleic Acids Research 12 (1): 387 (1984); five BLAST program sets, three designed for nucleotide sequence queries (BLASTN, BLASTX, and TBLASTX) and two designed for protein sequence queries (BLASTP and TBLASTN) Coulson, Trends in Biotechnology, 12: 76-80 (1994); Birren, et al., Genome Analysis, 1: 543-559 (1997)] BLAST X programs are commonly used in NCBI and other origins. ( BLAST Manual , Altschul, S., et al ., NCBI NLM NIH, Bethesda, MD 20894); Altschul, S., et al ., J. Mol. Biol ., 215: 403-410 (1990) The best known Smith Waterman algorithm can be used to confirm homology.

In general, parameters for polypeptide sequence comparison include: Algorithm: Needleman and Wunsch, J. Mol. Biol. 48: 443-453 (1970); Comparision Matrix: BLOSSUM 62 from Hentikoff and Hentikoff, Proc. Natl. Acad. Sci. USA 89: 10915-10919 (1992); Gap Penalty: 12; Gap Length Penalty: 4. A program that can be used for these parameters is one that is commonly used, such as the "gap" program of the genetics computer group ("GCG", Madison, Wisconsin). The parameter without penalty for the last gap is the default parameter for peptide comparison. In one embodiment, NCBI's BLASTP program does no compositional titration, with a predictive value of 10, a word size of 3, a BLOSUM62 matrix, a gap extension cost of 11, a final gap extension cost of 1, and blast extension (in bits). (X) Use the default parameter with an X dropoff value of 15 for gap alignment (in bits) and a final X dropoff value of 25 for gap alignment (in bits).

Provided herein is to reduce food intake, or to lose weight by administering at least one endoterin or an agonist thereof to a patient who wants to reduce food intake, weight loss (or maintenance), or both. It is a method for reducing (or maintaining) or both. These patients are generally at risk of developing or having a condition or disease that would benefit from reduced food intake, weight loss (or maintenance). Thus, one aspect provided herein is a method of weight loss, comprising administering an effective amount of an endothelin or endothelin agonist to a patient in need or need of weight loss. Further provided are methods of treating obesity comprising administering to a patient in need thereof an effective amount of an endoterin or endothelin agonist. Also provided are methods of preventing or treating metabolic disease in a patient in need of preventing or treating a metabolic disease, including administering an effective amount of an endoterin or an endothelin receptor agonist to a patient having or at risk for a metabolic disease. . In one embodiment, the metabolic disease is obesity, diabetes, insulin-resistant syndrome, metabolic syndrome, syndrome-X, or other disease resulting from excess calorie intake or uncontrolled food or energy metabolism. In one embodiment, the patient with a metabolic disease is in an overweight condition, overweight, requires weight loss or maintenance, or wants to inhibit weight gain in the future.

Endoterin can be endoterin-1, endoterin-2, endoterin-3, analogs, derivatives or functional fragments thereof, or combinations thereof. In certain embodiments, the endoterin is human endoterin-3 (SEQ ID NO: 10). In one embodiment, the endothelin or endothelin agonist acts through the ET _B receptor. In certain embodiments, the endothelin or endothelin agonist does not act through the ET _A receptor has a reduced affinity for ET _A receptors. In other embodiments, the endothelin or endothelin agonist preferably acts via the ET _B receptor. In one embodiment, a patient requiring or requiring treatment of a condition or disease that can be alleviated by reducing food intake by administering an effective dose of an endoterin or endothelin agonist to reduce food intake. It does not include patients receiving symptomatic endotherin or endothelin agonists.

Provided herein is a method for suppressing appetite or feeling of satiety, comprising administering an effective amount of an endoterin or endothelin agonist to a patient in need or need for suppressing appetite or increasing satiety.

In addition, the methods provided herein may further comprise administering additional compounds that act to increase satiety, reduce food intake, or maintain or reduce weight. Such additional compounds include, but are not necessarily limited to, exendin or agents thereof (see, eg, US Pat. No. 6,956,026; WO 99/25727; WO99 / 25728; WO99 / 07404); Amylin or agents thereof (see, eg, Pittner et al., J. Cell. Biochem. 55S: 19-28 (1994); US Pat. Nos. 5,686,411; 6,610,824; 6,410,511). ); Cholecystokinin (CCK) (see, eg , US Pat. No. 5,739,106; 5,270,302), oxygenntomodulin (see, eg, US Pat. No. 5,858,975), peptide YY (PYY) (US) Patent applications 2005/0176643; 2002/0141985) or leptin (ob protein) (see, eg , US Pat. No. 6,475,984; 6,399,745). Suitable amylin agents are for example [ ^25,28,29 Pro-]-human amylin (also called “pramlintide”) (eg, US Pat. Nos. 5,175,145; 5,814,600; 5,998,367). 6,114,304; 6,410,511; 6,608,029; 6,6610,824), salmon calcitonin and the compounds described in the following patents US20050197287, WO2006105345, WO2006083254, WO2006052608, and WO2005115437. All of which are incorporated by reference to the amylin family disclosed herein. The CCK used in one embodiment is a CCK octopeptide (CCK-8). Endothelin and endothelin agonists may be administered alone or in admixture with one or more such compounds. The method of administration can be performed concurrently, continuously or continuously with similar or other compounds.

Also included is the reduction of food intake, weight loss (or administering at least one endothelin or endothelin receptor agonist to a patient in need or need of reduced food intake, weight loss (or maintenance), or both. Maintenance) or the use of an endothelin or endothelin receptor agonist to perform both. Further provided are the use of endoterin or endoterin receptor agonists to prepare drugs suitable for patients in need of or needing reduced food intake, weight loss, or both.

In one aspect, the methods of the present invention relate to methods of reducing weight and / or reducing weight gain in an animal, and more particularly to methods of treating or alleviating obesity in a patient at risk or suffering from obesity. In one embodiment the method is effective to reduce the animal's weight to a measurable amount. In one embodiment, the weight loss of the animal can be measured within at least about 2 weeks, sometimes within about 4 weeks, preferably at least 6-8 weeks from the step of administering the compound.

In other embodiments, all of the methods disclosed herein reduce the weight of the patient by at least 1%, at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, or at least 50%. In further embodiments, all of the methods disclosed herein weigh a patient at least about 5 pounds or 2 kg, at least about 10 pounds or 5 kg, at least about 20 pounds or 10 kg, at least about 30 pounds or 15 kg, at least about 40 Pounds or 20 kg, at least about 50 pounds or 25 kg, at least about 75 pounds or 35 kg, at least about 100 pounds or 50 kg, at least about 125 pounds or 55 kg, at least about 150 pounds or 75 kg, at least about 175 pounds or Reduce by 80 kg, or at least about 200 pounds or 100 kg. In another embodiment, all of the methods disclosed herein can result in weight loss and can be about 40% or less, about 20% or less, about 10% or less, about 2% or less, about 1% or less of average weight loss. Or about 0% is caused by a decrease in mean body mass.

In another embodiment, the average food intake of the patient is reduced by at least 50%, 40%, 35%, 30%, 25%, compared to the average food intake of the patient not administered the endothelin or endothelin receptor agonist disclosed herein 20%, 15%, 10%, or 5% reduction. Average food intake rates can be ascertained for all defined time periods, eg, per meal, daily, weekly or monthly. In another embodiment, the daily average food intake of the patient is reduced by at least 2000 calories, at least 1500 calories, or at least 1000 calories compared to the food intake of the patient not administering the endothelin or endothelin receptor agonist disclosed herein. In another embodiment, the daily average food intake of the patient is 2000 calories, 1500 calories, 1000 calories, 750 calories, 500 calories, 250 compared to the food intake of the patient not administered the endothelin or endothelin receptor agonist disclosed herein. Calories or 100 calories are reduced. As used herein, calories refer to nutritional calories known as large calories or kilogram calories. Nutritional calorie is a unit that represents the heat- or energy-consuming value of a food that can produce one large energy calorie (1000 gram calories or 3.968 Btu) when the food is oxidized in the body.

The condition or disease can be treated or prevented by administering to the patient an effective amount of a compound of the invention for the amount or time necessary to achieve the desired result. The amount of endoterin or endoterin agonist that effectively alleviates the disease by reducing food intake or by losing weight (or maintaining) is a suitable benefit / risk ratio applicable to all medical treatments. Therefore, the dose should not have undesirable cardiovascular effects on the patient. Specific effective dose levels for all patients include the condition or condition to be treated and the severity of the condition or disease; Activity of the compound used; The specific composition employed; The age, body weight, health status, sex and diet of the patient; Dosing time; Route of administration, rate of excretion; Duration of treatment; And other activating agents used for mixed or concurrent treatment. When used by a physician, the composition will be presented in dosage unit form containing the amount of endoterin or endothelin agonist, with or without other food intake-reduction, plasma glucose-lowering agents, or tongue lipid-lowering agents. .

The term "effective amount" refers to the amount of a compound, alone or in combination, according to the methods disclosed herein, necessary for reduced food intake, weight loss (or maintenance) or both. In general, the daily effective food intake-reducing dose of the compound is administered in single or divided doses, from about 10 μg to about 5 mg / day, from about 10 μg to about 2 mg / day, from about 10 μg to about 1 mg / Days, or about 30 μg to about 500 μg / day. In another embodiment, the dosage range of endoterin or sarapotoxin is about 1 nmol / kg to 10 micromol / kg, in other embodiments about 1 nmol / kg to about 1000 nmol / kg, and in another embodiment about 10 nmol / kg to about 1000 nmol / kg, and in other embodiments from about 100 nmol / kg to 1000 nmol / kg. In certain embodiments, the dose of adenoregulin or demasceptin will be 5-10 times greater than the dose of endoterin and sarapotoxin. Thus, in one embodiment, the dose of adenoregulin or sarapotoxin is from about 5 nmol / kg to about 1 millimol / kg, in other embodiments from about 5 nmol / kg to about 10 micromol / kg, in another embodiment From about 50 nmol / kg to about 10 micromol / kg, in other embodiments from about 500 nmol / kg to about 10 micromol / kg. In one embodiment, these doses may be increased or decreased depending on the weight of the patient. In general, administration of an endothelin or endothelin receptor agonist can begin at any time where inhibition of food intake, weight loss or weight maintenance is necessary, for example obesity, diabetes, insulin-resistant syndrome or other conditions or diseases disclosed herein. It can begin immediately after the first sign of symptoms or after they have been diagnosed. Administration can be by injection, eg, subcutaneously, intramuscularly, intraperitoneally or intravenously. Oral active compounds can be administered orally, but dosages are generally increased by 5-10 fold. In one embodiment, the compound or composition is administered daily, weekly or monthly. Dosage may be reduced until the weight of the individual is reduced until a base dose is achieved or no further dose is required. In another embodiment, the compound is administered immediately before the meal. In another embodiment, the compound is administered at the start of a meal.

In one aspect, the effective amount is a dose sufficient to reduce food intake, reduce weight, or cause both without noticeable or significant effects on the cardiovascular system. The effect on the cardiovascular system may be an increase or decrease in blood pressure, eg, systolic, diastolic, or both; Increase in total peripheral vascular resistance; Or increased pulmonary vascular resistance. In this embodiment, the dose is an amount effective to cause reduced food intake, weight loss, or both, and does not change the mean systolic blood pressure, mean diastolic blood pressure, or both above 5 mm Hg. In another embodiment, the dose is an amount effective to cause reduced food intake, weight loss, or both, and does not change the mean systemic systolic blood pressure, mean systemic diastolic blood pressure, or both above 5 mm Hg. In one embodiment, the dose is an amount effective to cause reduced food intake, weight loss, or both, and does not change the average systolic blood pressure above 139 mm Hg and the average diastolic blood pressure above 89 mm Hg. In another embodiment, the dose is an amount effective to cause reduced food intake, weight loss, or both, and does not change the ratio of systolic blood pressure to diastolic blood pressure above 139 to 89 mm Hg.

Data obtained from cell culture assays and animal experiments can be used to adjust the dose range for use in humans or other animals. In general, the dosage of such compounds is within a range of circulating concentrations that include ED ₅₀ , which is non-toxic or slightly toxic. In particular, the methods of the present invention include ET agents to avoid dangerous vascular action, especially in cardiovascular systems. Dosages may vary within the range depending on the dosage form employed and the route administered. For all compositions used in the methods disclosed herein, the therapeutically effective dosage can be determined first from the results of the cell culture assay. Dosages may be determined in animal models to reach circulating plasma concentrations including EC ₅₀ (ie the concentration of tested compound reaching half of the maximum effective response) as determined in cell culture. This information can be used to more accurately determine the effective dose for humans. Plasma levels can be determined, for example, by high performance liquid chromatography or antibody assays.

The optimal formulation and optimal method of administration for applying a compound of the present invention to a patient depends on factors known in the art such as the specific condition or disease, the desired effect, the type of patient and the patient's tendency.

Compounds effective in the methods of the invention may typically be presented in the form of formulations suitable for parenteral (including intravenous, intraperitoneal, intramuscular and subcutaneous) or intranasal or oral administration. In one embodiment, the route of administration is subcutaneous or intramuscular. In certain instances, endothelin or endothelin receptor agonists and exendin-4, exendin agonists, amylin, amylin agonists, at least one food intake reducing agent such as PYY, CKK, or leptin, plasma glucose-lowering agents or plasma lipids It will be convenient to provide the lowering agent in a single composition or as a solution for administration together. In other cases, it may be further advantageous to administer at least one additional agent independent of the endothelin or endothelin agonist. Appropriate dosage forms will preferably be determined by a physician skilled in the art for each patient. Suitable pharmaceutically acceptable carriers and formulations thereof are described in standard formulation literature. For example , Remington: The Science and Practice of Pharmacy (21st Ed. Lipincott, Williams, & Wilkins 2005).

As disclosed herein, endothelin and endothelin agonists can be used in combination with any conventional method or composition suitable for reducing food intake or promoting or maintaining weight loss. Such methods and means include, but are not necessarily limited to, diet, exercise, orlistat, sibutramine, obesity surgery, and allopathic suppression aids.

Therapeutic compositions of the present invention comprise an effective amount of a compound formulated with one or more therapeutically suitable excipients. As used herein, the term “therapeutically suitable excipient” refers to nontoxic solid, semisolid or liquid filters, diluents, encapsulating materials or auxiliary forms of all forms. Examples of therapeutically acceptable excipients include sugar; Cellulose and its derivatives; oil; Glycols; solution; Buffers, pigments, secretions, coatings, sweeteners, flavorings, fragrances, and the like. These therapeutic compositions can be administered parenterally, in a bath, orally, rectally or intraperitoneally.

Liquid dosage forms for oral administration of the compositions of the invention include formulations such as emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to this compound, liquid dosage forms can include diluents and / or solubilizers or emulsifiers. In comparison to inert diluents, oral compositions may include humectants, emulsifiers, sweeteners, flavors and fragrances. Injectable formulations of the compositions of the present invention include sterile, injectable liquid and oily solutions, suspensions or emulsions, all of which may optionally be formulated with parenterally suitable diluents, dispersants, wetting agents or suspending agents. These injection formulations may be formulated into sterile formulations which are sterilized by filtration through a bacterina-conserving filter or dissolved or dispersed in an injection medium.

Compounds useful in the methods of the invention may be provided as parenteral compositions for injection or infusion. For example they can be suspended by inert oils, suitably sesame oil, peanut oil, vegetable oils such as olive oil or other acceptable carriers. These compositions may be sterilized or sterile filtered by conventional sterilization techniques. The composition may comprise pharmaceutically acceptable auxiliary ingredients necessary for a suitable physiological condition, such as pH buffers. Useful buffers include, for example, sodium acetate / acetic acid buffers. If a depot, sustained or “depot” slow release formulation form is used, a therapeutically effective amount of the formulation may be delivered through the blood vessel for hours or days following transdermal injection or delivery.

Preferred isotonic osmotic pressures include other therapeutically acceptable agents such as sodium chloride or dextrose, boric acid, sodium tartrate, propylene glycol, polyols (such as mannitol and sorbitol), or other inorganic or organic solutions. Can be realized. Sodium chloride is particularly preferred as a buffer containing sodium ions.

Compounds useful in the methods and therapies disclosed herein may be in the form of an acid (-OH) or in the form of an N terminal amidated (-NH ₂ ). In addition, the compounds may be formulated as pharmaceutically acceptable salts (eg, acid addition salts) and / or complexes thereof. Pharmaceutically acceptable salts are non-toxic salts at the concentrations when administered. Formulations of such salts may facilitate pharmaceutical use by changing the physiological-chemical properties of the composition without inhibiting the composition from its physiological effects. Examples of effective changes in physiological properties include lowering the melting point to facilitate mucosal administration and increasing solubility to promote the administration of high concentrations of the drug.

Pharmaceutically acceptable salts include sulfate, hydrochloride, phosphate, sulfamate, acetate, citrate, lactate, tartrate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, cyclo Acid addition salts such as those including hexylsulfate and quinate. Pharmaceutically acceptable salts include hydrochloric acid, sulfuric acid, phosphoric acid, sulfamic acid, acetic acid, citric acid, lactic acid, tartaric acid, malonic acid, methanesulphonic acid, ethanesulphonic acid, benzenesulphonic acid , can be obtained from acids such as p-toluenesulphonic acid, cyclohexylsulfonic acid and quinic acid. Such salts are reacted with the free acid or base form product using one or more equivalent suitable bases or acids, for example in a solvent or medium in which the salt is insoluble, or in a solvent such as water, followed by vacuum or freeze-drying. Or by removing the salts by ion exchange exchanging remaining salts with other ions in a suitable ion exchange resin.

In addition, carriers or excipients may be used to facilitate the administration of the compounds. Examples of carriers and excipients include various sugars such as calcium carbonate, calcium phosphate, lactate, glucose or sucrose, or starch forms, cellulose derivatives, gelatin, vegetable oils, polyethylene glycols and physiologically acceptable solvents. The composition or pharmaceutical composition may be administered by intravenous, intraperitoneal, subcutaneous and intramuscular, oral, topical, mucous pain or alveolar inhalation.

If desired, the solution of the composition may be concentrated by a thickening agent such as methyl cellulose. It may also be prepared in an emulsified form, such as oil-in-water or water-in-oil. A wide range of pharmaceutically acceptable emulsifiers include, for example, acacia powder, nonionic surfactants (such as Tween), or ionic surfactants (such as alkali polyester alcohol sulfates or sulfonates, for example Triton). It may include.

Compositions useful in the present invention are prepared by mixing the components in accordance with generally acceptable methods. For example, the selected ingredients are simply mixed in a mixer or general equipment for producing a concentrated mixture, which are optimized for final concentration and viscosity by adding water or thickener, and adding a buffer to adjust pH. Or solutes may be added to control the osmotic pressure.

In order to understand the method of the present invention, the following examples are included. The following experiments relate to the method of the present invention and do not specifically limit the present invention, and modifications of the present invention which fall within the scope of the presently known or later occurring ones are described herein and as claimed in the following claims. It is considered to be within the scope of the present invention.

Example 1

Endoterin  And Endoterin  Reduction of food intake by agents

Experimental Materials and Methods: NIH / Swiss mice were starved approximately 17 hours prior to the start of the experiment. Peptides were injected intraperitoneally (ip) at zero time at doses as indicated. All mice were intraperitoneally injected (200 μl) with mediators or indicated doses of compound and immediately given pre-weighed pellets. The pellets were left to check the amount of food eaten and weighed at intervals of 30 minutes, 1 hour and 2 hours. Each point represents 4 mice receiving endothelin or endothelin agonist or 3 mice receiving medium. The compounds used in the experiments are shown in Table 5 below.

compound name order Endoterin 1 c (CSCSSLMDKECVYFCHLDIIW) -OH SEQ ID NO: 1 Endoterin-3 c (CTCFTYKDKECVYYCHLDIIW) -OH SEQ ID NO: 10 Endoterin-2 c (CSCSSWLDKECVYFCHLDIIW) -OH SEQ ID NO: 7 [Succinyl (Glu ⁹ , Ala ¹¹ , Ala ¹⁵ ] -Endoterin-1- (8-21)] (IRL1620) Succinyl-DEEAVYFAHLDIIW-OH SEQ ID NO: 19 Sarapotoxin S6c [ Atractaspis engaddensis ] c (CTCNDMTDEECLNFCHQDVIW) -OH SEQ ID NO: 17

The food intake is expressed in each time point, and the effect of the test sample ingested is expressed as% change for the medium using the following equation.

% Basal = -100 * [1-(food intake of treatment group / food intake of vehicle group)].

Significant test sample effects were confirmed by ANOVA (p <0.05). When significant differences appeared, Dunnett's test was used to compare the mean of the experimental group to the control mean. One-way ANOVA using Dunnett's post test was performed using GraphPad Prism® version 3.01 for Windows (GraphPad Software, San Diego, California USA).

RESULTS: There was a dose-dependent decrease in food intake at all time points after peripheral administration of ET-2, ET-3 and sarapotoxin S6c as well as ET-1. 1, 2 and 3. The dose response to the reduced food intake at 30 minutes showed a similar effect as tested for the agent, but the reduced food intake at 60 minutes was moderate. See FIGS. 2 and 3. IRL1620 reduced food intake at 30 and 60 minutes, but had no effect at 120 minutes, due to the much faster removal / inactivation of the molecule. These data demonstrate that ET-1, ET-2, and ET-3 are potent and fully effective anorexigens in hungry mice without exhibiting observable side effects. The effect of ET-3 on reducing food intake suggests that the appetite suppressing effect is induced by the ET _B receptor and that ET-3 binds weakly to ET _A.

Example 2

Weight Loss by Endothelin and Endothelin Agonists in DIO Mice

Experimental Materials and Methods: Dietary induced obesity (DIO) mice were used. Obesity was induced by dieting pellets of high fat diet (58% calories, # D12331, Research Diets, New Brunswick, NJ) in 4 week old mice (20) for 6 weeks prior to treatment (4 weeks for long-term study). The mice were then maintained in powder form during the treatment, unless otherwise indicated. All animals were raised under 21-23 ° C. conditions with a 12-hour: 12-hour light-dark cycle, and were given food at random before and after treatment. Mediators, ET-1 [SEQ ID NO: 1] (20 nmol / kg / d), or adenoregulin [SEQ ID NO: 33] (300 nmol / kg / d) were administered to DIO mice with an Alzet ^® sc osmotic pump. Mice were fed high fat diet pellets and recorded weekly weight and food intake.

Body weight (BWt) at each time point is expressed as% change to mediator treatment and represents the effect on BWt of the test sample. For the treatment group:

% BWt loss = 100 * [(Average BWt change of media treatment group)-(Average BWt change of treatment group) / Average BWt on day 0]

For each:

% BWt loss = 100 * [(mean change of BWt of the medium treatment group)-(change of BWt of the treatment group) / mean BWt of day 0].

Test samples showing significant effects were identified by ANOVA (p <0.05). When significant differences appeared, Dunnett's test was used to compare the mean of the experimental group to the control mean. One-way ANOVA using Dunnett's post test was performed using GraphPad Prism ^® version 3.01 for Windows (GraphPad Software, San Diego, California USA).

RESULTS: DIO mice continuously infused with ET _B agonist, adenoregulin for 2-4 weeks, not only maintained the weight loss effect (FIG. 5) but also decreased body fat (FIG. 4). As a result, adenoregulin, which inhibited food intake in rodents, was much less effective than showing antibiotic effects. Similarly, continuously injected ET-1 also showed weight loss in DIO mice compared to the control (FIG. 6). This effect was maintained without observable symptoms. Taken together, this study demonstrates that ET-1 and ET _B agonists can reduce food intake and reduce body weight and body fat.

All publications and patents cited herein are hereby incorporated by reference, and each publication or patent is expressly incorporated by reference in its entirety. Although the preceding invention has been described in detail by way of explanation and illustration for purposes of clarity of understanding, it will not be left to the gist of the invention or the appended claims, even if a person of ordinary skill in the art of the present invention changes and changes slightly. It will be readily appreciated.

                         SEQUENCE LISTING <110> Amylin Pharmaceuticals, Inc        Young, Andrew        Pittner, Richard   <120> ENDOTHELIN AND ENDOTHELIN RECEPTOR AGONISTS IN THE TREATMENT OF        METABOLIC DISEASES <130> 1101WO1 <150> US 60 / 785,447 <151> 2006-03-23 <160> 64 <170> PatentIn version 3.3 <210> 1 <211> 21 <212> PRT <213> Homo sapiens <400> 1 Cys Ser Cys Ser Ser Leu Met Asp Lys Glu Cys Val Tyr Phe Cys His 1 5 10 15 Leu Asp Ile Ile Trp             20 <210> 2 <211> 21 <212> PRT <213> Felis catus <400> 2 Cys Ser Cys Ser Ser Leu Leu Asp Lys Glu Cys Val Tyr Phe Cys His 1 5 10 15 Leu Asp Ile Ile Trp             20 <210> 3 <211> 21 <212> PRT <213> Oncorhynchus keta <400> 3 Cys Ser Cys Ala Thr Phe Leu Asp Lys Glu Cys Val Tyr Phe Cys His 1 5 10 15 Leu Asp Ile Ile Trp             20 <210> 4 <211> 21 <212> PRT <213> Danio rerio <400> 4 Cys Ser Cys Ala Ser Phe Leu Asp Lys Glu Cys Val Tyr Phe Cys His 1 5 10 15 Leu Asp Ile Ile Trp             20 <210> 5 <211> 21 <212> PRT <213> Gallus gallus <400> 5 Cys Ser Cys Ser Ser Leu Leu Asp Glu Glu Cys Val Tyr Phe Cys His 1 5 10 15 Leu Asp Ile Ile Trp             20 <210> 6 <211> 21 <212> PRT <213> Monodelphis domestica <400> 6 Cys Ser Cys Ser Ser Leu Leu Asp Lys Glu Cys Val Tyr Phe Cys Leu 1 5 10 15 Leu Asp Ile Ile Trp             20 <210> 7 <211> 21 <212> PRT <213> Homo sapiens <400> 7 Cys Ser Cys Ser Ser Trp Leu Asp Lys Glu Cys Val Tyr Phe Cys His 1 5 10 15 Leu Asp Ile Ile Trp             20 <210> 8 <211> 21 <212> PRT <213> Rattus norvegicus <400> 8 Cys Ser Cys Asn Ser Trp Leu Asp Lys Glu Cys Val Tyr Phe Cys His 1 5 10 15 Leu Asp Ile Ile Trp             20 <210> 9 <211> 21 <212> PRT <213> Gallus gallus <400> 9 Cys Ser Cys Asn Ser Trp Leu Asp Lys Glu Cys Ile Tyr Phe Cys His 1 5 10 15 Leu Asp Ile Ile Trp             20 <210> 10 <211> 21 <212> PRT <213> Homo sapiens <400> 10 Cys Thr Cys Phe Thr Tyr Lys Asp Lys Glu Cys Val Tyr Tyr Cys His 1 5 10 15 Leu Asp Ile Ile Trp             20 <210> 11 <211> 21 <212> PRT <213> Monodelphis domestica <400> 11 Cys Thr Cys Tyr Thr Tyr Lys Asp Lys Glu Cys Val Tyr Tyr Cys His 1 5 10 15 Leu Asp Ile Ile Trp             20 <210> 12 <211> 21 <212> PRT <213> Bos taurus <400> 12 Cys Thr Cys Phe Thr Tyr Lys Asp Arg Glu Cys Val Tyr Tyr Cys His 1 5 10 15 Leu Asp Ile Ile Trp             20 <210> 13 <211> 21 <212> PRT <213> Danio rerio <400> 13 Cys Thr Cys Tyr Ser Tyr Lys Asp Lys Glu Cys Val Tyr Tyr Cys His 1 5 10 15 Leu Asp Ile Ile Trp             20 <210> 14 <211> 21 <212> PRT <213> Artificial <220> <223> Synthetic Endothelin-1 analog <400> 14 Ala Ser Ala Ser Ser Leu Met Asp Lys Glu Ala Val Tyr Phe Ala His 1 5 10 15 Leu Asp Ile Ile Trp             20 <210> 15 <211> 21 <212> PRT <213> Atractaspis engadensisan <400> 15 Cys Ser Cys Lys Asp Met Thr Asp Lys Glu Cys Leu Asn Phe Cys His 1 5 10 15 Gln Asp Val Ile Trp             20 <210> 16 <211> 21 <212> PRT <213> Atractaspic engadensisan <400> 16 Cys Ser Cys Lys Asp Met Thr Asp Lys Glu Cys Leu Tyr Phe Cys His 1 5 10 15 Gln Asp Val Ile Trp             20 <210> 17 <211> 21 <212> PRT <213> Atractaspic engadensisan <400> 17 Cys Thr Cys Asn Asp Met Thr Asp Glu Glu Cys Leu Asn Phe Cys His 1 5 10 15 Gln Asp Val Ile Trp             20 <210> 18 <211> 21 <212> PRT <213> Atractaspic engadensisan <400> 18 Cys Thr Cys Asn Asp Met Thr Asp Lys Glu Cys Leu Tyr Phe Cys His 1 5 10 15 Glu Asp Ile Ile Trp             20 <210> 19 <211> 14 <212> PRT <213> Artificial <220> <223> modified endothelin-1 fragment <220> <221> MOD_RES (222) (1) .. (1) <223> succinyl <400> 19 Asp Glu Glu Ala Val Tyr Phe Ala His Leu Asp Ile Ile Trp 1 5 10 <210> 20 <211> 81 <212> PRT <213> Phyllomedusa bicolor <220> <221> MOD_RES <222> (81) .. (81) <223> OPTIONAL AMIDATION <400> 20 Met Ala Phe Leu Lys Lys Ser Leu Phe Leu Val Leu Phe Leu Gly Leu 1 5 10 15 Val Ser Leu Ser Ile Cys Glu Glu Glu Lys Arg Glu Asn Glu Asp Glu             20 25 30 Glu Glu Gln Glu Asp Asp Glu Gln Ser Glu Met Lys Arg Gly Leu Trp         35 40 45 Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala Ala Ala Lys     50 55 60 Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala Val Gly Glu 65 70 75 80 Gln      <210> 21 <211> 78 <212> PRT <213> Phyllomedusa bicolor <220> <221> MOD_RES <222> (78) .. (78) <223> OPTIONAL AMIDATION <400> 21 Met Asp Ile Leu Lys Lys Ser Leu Phe Leu Val Leu Phe Leu Gly Leu 1 5 10 15 Val Ser Leu Ser Ile Cys Glu Glu Glu Lys Arg Glu Asn Glu Asp Glu             20 25 30 Glu Lys Gln Asp Asp Glu Gln Ser Glu Met Lys Arg Ala Met Trp Lys         35 40 45 Asp Val Leu Lys Lys Ile Gly Thr Val Ala Leu His Ala Gly Lys Ala     50 55 60 Ala Leu Gly Ala Val Ala Asp Thr Ile Ser Gln Gly Glu Gln 65 70 75 <210> 22 <211> 77 <212> PRT <213> Phyllomedusa bicolor <220> <221> MOD_RES (222) (77) .. (77) <223> OPTIONAL AMIDATION <400> 22 Met Ala Phe Leu Lys Lys Ser Leu Phe Leu Val Leu Phe Leu Gly Leu 1 5 10 15 Val Ser Leu Ser Ile Cys Glu Glu Glu Lys Arg Glu Asn Glu Asp Glu             20 25 30 Glu Glu Gln Glu Asp Asp Glu Gln Ser Glu Met Lys Arg Gly Leu Trp         35 40 45 Ser Lys Ile Lys Glu Ala Gly Lys Ala Ala Leu Thr Ala Ala Gly Lys     50 55 60 Ala Ala Leu Gly Ala Val Ser Asp Ala Val Gly Glu Gln 65 70 75 <210> 23 <211> 76 <212> PRT <213> Phyllomedusa bicolor <220> <221> MOD_RES <222> (76) .. (76) <223> OPTIONAL AMIDATION <400> 23 Met Ala Phe Leu Lys Lys Ser Leu Phe Leu Val Leu Phe Leu Gly Leu 1 5 10 15 Val Ser Leu Ser Ile Cys Glu Glu Glu Lys Arg Glu Asn Lys Asp Glu             20 25 30 Ile Glu Gln Glu Asp Asp Glu Gln Ser Glu Glu Lys Arg Ala Leu Trp         35 40 45 Lys Asp Ile Leu Lys Asn Val Gly Lys Ala Ala Gly Lys Ala Val Leu     50 55 60 Asn Thr Val Thr Asp Met Val Asn Gln Gly Glu Gln 65 70 75 <210> 24 <211> 81 <212> PRT <213> Phyllomedusa bicolor <220> <221> MOD_RES <222> (81) .. (81) <223> OPTIONAL AMIDATION <400> 24 Met Ala Ser Leu Lys Lys Ser Leu Phe Leu Val Leu Phe Leu Gly Leu 1 5 10 15 Val Ser Leu Ser Ile Cys Glu Glu Glu Lys Arg Glu Asn Glu Asp Glu             20 25 30 Glu Glu Gln Glu Asp Asp Glu Gln Ser Glu Met Lys Arg Gly Leu Trp         35 40 45 Ser Asn Ile Lys Thr Ala Gly Lys Glu Ala Ala Lys Ala Ala Leu Lys     50 55 60 Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Thr Asp Ala Val Gly Glu 65 70 75 80 Gln      <210> 25 <211> 72 <212> PRT <213> Phyllomedusa bicolor <220> <221> MOD_RES (222) (72) .. (72) <223> OPTIONAL AMIDATION <400> 25 Met Asp Ile Leu Lys Lys Ser Leu Phe Leu Val Leu Phe Leu Gly Leu 1 5 10 15 Val Ser Leu Ser Ile Cys Glu Glu Glu Lys Arg Glu Asn Glu Asp Glu             20 25 30 Glu Lys Gln Asp Asp Glu Gln Ser Glu Met Lys Arg Ala Met Trp Lys         35 40 45 Asp Val Leu Lys Lys Ile Ala Gly Lys Ala Ala Leu Gly Ala Val Ala     50 55 60 Asp Thr Ile Ser Gln Gly Glu Gln 65 70 <210> 26 <211> 72 <212> PRT <213> Phyllomedusa bicolor <220> <221> MOD_RES (222) (72) .. (72) <223> OPTIONAL AMIDATION <400> 26 Met Ala Phe Leu Lys Lys Ser Leu Phe Leu Val Leu Phe Leu Gly Leu 1 5 10 15 Val Ser Leu Ser Val Cys Glu Glu Glu Lys Arg Glu Asn Glu Asp Glu             20 25 30 Met Glu Gln Glu Asp Asp Glu Gln Ser Glu Glu Lys Arg Ala Leu Trp         35 40 45 Lys Asp Ile Leu Lys Asn Ala Gly Lys Ala Ila Leu Asn Glu Ile Asn     50 55 60 Gln Leu Val Asn Gln Gly Glu Leu 65 70 <210> 27 <211> 74 <212> PRT <213> Phyllomedusa bicolor <220> <221> MOD_RES <222> (74) .. (74) <223> OPTIONAL AMIDATION <400> 27 Met Ala Phe Leu Lys Lys Ser Val Phe Leu Val Leu Phe Leu Gly Leu 1 5 10 15 Val Ser Leu Ser Ile Cys Glu Glu Glu Lys Arg Glu Glu Glu Asn Glu             20 25 30 Glu Lys Gln Glu Asp Asp Glu Gln Ser Glu Glu Lys Arg Ala Leu Trp         35 40 45 Lys Asn Met Leu Lys Gly Ile Gly Lys Leu Ala Gly Gln Ala Ala Leu     50 55 60 Gly Ala Val Lys Thr Leu Val Gly Ala Glu 65 70 <210> 28 <211> 77 <212> PRT <213> Phyllomedusa bicolor <220> <221> MOD_RES (222) (77) .. (77) <223> OPTIONAL AMIDATION <400> 28 Met Ala Phe Leu Lys Lys Ser Leu Phe Leu Val Leu Phe Leu Gly Leu 1 5 10 15 Val Pro Leu Ser Leu Cys Glu Ser Glu Lys Arg Glu Gly Glu Asn Glu             20 25 30 Glu Glu Gln Glu Asp Asp Gln Ser Glu Glu Lys Arg Ser Leu Gly Ser         35 40 45 Phe Leu Lys Gly Val Gly Thr Thr Leu Ala Ser Val Gly Lys Val Val     50 55 60 Ser Asp Gln Phe Gly Lys Leu Leu Gln Ala Gly Gln Gly 65 70 75 <210> 29 <211> 26 <212> PRT <213> Phyllomedusa bicolor <220> <221> MOD_RES (222) (26) .. (26) <223> OPTIONAL AMIDATION <400> 29 Gly Leu Trp Asn Lys Ile Lys Glu Ala Ala Ser Lys Ala Ala Gly Lys 1 5 10 15 Ala Ala Leu Gly Phe Val Asn Glu Met Val             20 25 <210> 30 <211> 27 <212> PRT <213> Phyllomedusa bicolor <220> <221> MOD_RES <222> (27) .. (27) <223> OPTIONAL AMIDATION <400> 30 Asp Val Leu Lys Lys Ile Gly Thr Val Ala Leu His Ala Gly Lys Ala 1 5 10 15 Ala Leu Gly Ala Val Ala Asp Thr Ile Ser Gln             20 25 <210> 31 <211> 21 <212> PRT <213> Atractaspis microlepidota <400> 31 Cys Ser Cys Asn Asp Met Asn Asp Lys Glu Cys Met Tyr Phe Cys His 1 5 10 15 Gln Asp Val Ile Trp             20 <210> 32 <211> 21 <212> PRT <213> Artificial <220> <223> Synthesized sarafotoxin <400> 32 Cys Ser Cys Lys Asp Met Ser Asp Lys Glu Cys Leu Asn Phe Cys His 1 5 10 15 Gln Asp Val Ile Trp             20 <210> 33 <211> 33 <212> PRT <213> Phyllomedusa bicolor <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 33 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Val      <210> 34 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 34 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Val      <210> 35 <211> 32 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (32) .. (32) <223> OPTIONAL AMIDATION <400> 35 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 <210> 36 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 36 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Phe      <210> 37 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (31) .. (31) <223> OPTIONAL AMIDATION <400> 37 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Leu      <210> 38 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 38 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Ile      <210> 39 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 39 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Tyr      <210> 40 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 40 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Trp      <210> 41 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 41 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Ala      <210> 42 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 42 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Lys      <210> 43 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 43 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Pro      <210> 44 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 44 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Arg      <210> 45 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 45 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 His      <210> 46 <211> 32 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (32) .. (32) <223> OPTIONAL AMIDATION <400> 46 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 <210> 47 <211> 36 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES <222> (36) .. (36) <223> OPTIONAL AMIDATION <400> 47 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Val Gly Glu Gln         35 <210> 48 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 48 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Ile Lys Ala Ala Leu Ile Ala Val Ser Glu Ala             20 25 30 Val      <210> 49 <211> 32 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (32) .. (32) <223> OPTIONAL AMIDATION <400> 49 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 <210> 50 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 50 Gly Leu Trp Ser Ala Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Val      <210> 51 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 51 Gly Leu Ala Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Val      <210> 52 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 52 Ala Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Val      <210> 53 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 53 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Ala Ser Glu Ala             20 25 30 Val      <210> 54 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 54 Gly Ala Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Val      <210> 55 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 55 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Ala      <210> 56 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 56 Gly Leu Trp Ala Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Val      <210> 57 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 57 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ala Glu Ala             20 25 30 Val      <210> 58 <211> 33 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (33) .. (33) <223> OPTIONAL AMIDATION <400> 58 Gly Leu Trp Ser Lys Ile Lys Glu Val Gly Lys Glu Ala Ala Lys Ala 1 5 10 15 Ala Ala Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Ala Ala             20 25 30 Val      <210> 59 <211> 34 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (34) .. (34) <223> OPTIONAL AMIDATION <400> 59 Ala Leu Trp Lys Thr Met Leu Lys Lys Leu Gly Thr Met Ala Leu His 1 5 10 15 Ala Gly Lys Ala Ala Leu Gly Ala Ala Ala Asp Thr Ile Ser Gln Gly             20 25 30 Thr gln          <210> 60 <211> 31 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (31) .. (31) <223> AMIDATION <400> 60 Ala Leu Trp Lys Thr Leu Leu Lys Lys Val Gly Lys Val Ala Gly Lys 1 5 10 15 Ala Val Leu Asn Ala Val Thr Asn Met Ala Asn Gln Asn Glu Gln             20 25 30 <210> 61 <211> 27 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES <222> (27) .. (27) <223> OPTIONAL AMIDATION <400> 61 Ala Leu Trp Met Thr Leu Leu Lys Lys Val Leu Lys Ala Ala Ala Lys 1 5 10 15 Ala Leu Asn Ala Val Leu Val Gly Ala Asn Ala             20 25 <210> 62 <211> 13 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES (222) (13) .. (13) <223> OPTIONAL AMIDATION <400> 62 Ala Leu Trp Lys Thr Leu Leu Lys Lys Val Leu Lys Ala 1 5 10 <210> 63 <211> 36 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES <222> (36) .. (36) <223> OPTIONAL AMIDATION <400> 63 Gly Met Trp Ser Lys Ile Lys Asn Ala Gly Lys Ala Ala Ala Lys Ala 1 5 10 15 Ser Lys Lys Ala Ala Gly Lys Ala Ala Leu Gly Ala Val Ser Glu Ala             20 25 30 Leu Gly Glu Gln         35 <210> 64 <211> 27 <212> PRT <213> Artificial <220> <223> Synthesized adenoregulin <220> <221> MOD_RES <222> (27) .. (27) <223> OPTIONAL AMIDATION <400> 64 Gly Met Trp Gly Ser Leu Leu Lys Gly Val Ala Thr Val Val Lys His 1 5 10 15 Val Leu Pro His Ala Leu Ser Ser Gln Gln Ser             20 25  

Claims (74)

Reducing food intake in patients who want or need to reduce food intake, comprising administering to the patient who wants or needs to reduce food intake, an amount of endothelin or an endothelin agonist effective in reducing food intake. Way. A method of weight loss in a patient wishing or in need of weight loss, comprising administering to the patient wanting or in need of weight loss an amount of an endoterin or an endothelin agonist effective for weight loss. A method of treating obesity in a patient in need or need of treatment of obesity, comprising administering to a patient in need or need of treatment for obesity, an effective amount of an endoterin or an endothelin agonist for the treatment of obesity. Metabolism in a patient in need or need of treatment of a metabolic disease, comprising administering to the patient a desire or need for treatment of a metabolic disease, a dose of an endoterin or an endothelin agonist effective to treat the metabolic disease How to prevent or treat a disease. The method of claim 4, wherein the metabolic disease is obesity, diabetes, insulin-resistant syndrome, syndrome-X, or other hypertrophic disease. 6. The patient according to any one of claims 1 to 5, wherein the patient gains weight in the future in association with a disease or condition such as obesity, overweight, a patient in need of weight loss or weight gain caused by drugs. A patient in need of suppression. 7. The method according to any one of claims 1 to 6, wherein said endoterin is at least one of endoterin-1, endoterin-2, or endoterin-3. 8. The method of claim 7, wherein said endoterin is a sequence having at least 75% sequence homology with endoterin-1 or SEQ ID NO: 1. 8. The method of claim 7, wherein said endoterin is a sequence having at least 75% sequence homology with endoterin-2 or SEQ ID NO: 7. 8. A method according to claim 7, wherein said endoterin is a sequence having at least 75% sequence homology with endoterin-3 or SEQ ID NO: 10. The method according to any one of claims 1 to 6, wherein the endothelin agonist is an analog of SEQ ID NO: 1, SEQ ID NO: 7, or SEQ ID NO: 10 which does not include substitutions, deletions or additions of at least 5 amino acids. How to. 12. The method of claim 11, wherein the analogue does not comprise a substitution of at least 5 amino acids. 7. The method according to any one of claims 1 to 6, wherein said endoterin is endoterin-1 selected from at least one of the groups consisting of SEQ ID NOs: 1-6. 7. The method according to any one of claims 1 to 6, wherein said endoterin is endoterin-2 selected from at least one of the group consisting of SEQ ID NOs: 7-9. 7. The method according to any one of claims 1 to 6, wherein said endoterin is endoterin-3 selected from at least one of the groups consisting of SEQ ID NOs: 10-13. 7. The method of any one of claims 1 to 6, wherein the endothelin agonist is an endothelin analog that does not contain at least 35 amino acids. 17. The method of any of claims 7-16, wherein the endothelin agonist comprises a C terminus having the amino acid sequence FCHLDIIW. The method of claim 7, wherein the endothelin agonist comprises a C terminus having the amino acid sequence FAHLDIIW. 7. The endothelin agonist according to any one of claims 1 to 6, wherein the endothelin agonist is N-cis-2,6-dimethylpiperidinocarbonyl-L-γ-methylleucine-D-1-methoxycarbonyltrip Topanyl-Dnorleucine. 7. The method of any one of claims 1 to 6, wherein said endothelin agonist is [Ala ^{1, 3, 11, 15} ] endothelin-1 (SEQ ID NO: 14). The method according to any one of claims 1 to 6, wherein the endothelin agonist is a fragment of endothelin-1, endothelin-2, endothelin-3, and the fragment binds to and activates an ET _A or ET _B receptor. Characterized in that the method. 22. The method of claim 21, wherein said fragment does not comprise at least seven amino acid deletions. 7. The method of any one of claims 1 to 6, wherein the endothelin agonist does not comprise at least 10 amino acid substitutions, additions or deletions as compared to any one of SEQ ID NOs: 1, 7 and 10. 23. The method of claim 21 or 22, wherein the endothelin agonist is (N-succinyl- [Glu ⁹ , Ala ^11,15 ] ^-endoterin -1 fragment 8-21. 24. The method of claim 22 or 23, wherein said endothelin agonist is (N-acetyl- [Ala ^11,15 ] ^-endoterin -1 fragment 6-21. 7. The method of any one of claims 1 to 6, wherein the endothelin agonist is sarafotoxin. 27. The method of claim 26, wherein said sarapotoxin has an amino acid sequence selected from the group consisting of SEQ ID NOs: 15, 16, 17, 18, 31 and 32. 27. The method of claim 26, wherein the sarapotoxin has an amino acid sequence having at least 75% sequence homology with SEQ ID NOs: 15, 16, 17, 18, 31, and 32. 29. The method of any one of claims 26-28, wherein the sarapotoxin comprises a C terminus having the amino acid sequence FCHQDVIW. 7. The method of any one of claims 1 to 6, wherein the endothelin agonist is adenoregulin. 31. The method of claim 30, wherein said adenoregulin has an amino acid sequence selected from the group comprising SEQ ID NOs: 20-30. 31. The method of claim 30, wherein said adenoregulin has an amino acid sequence having at least 75% sequence homology to any one of SEQ ID NOs: 20-30. 33. The method of any one of claims 1 to 32, wherein said endothelin or endothelin agonist is an ET _B receptor agonist but not an ET _A receptor agonist. 33. The method of any one of claims 1 to 32, wherein the endothelin or endothelin agonist binds to and activates the ET _B receptor as compared to the ET _A receptor. 33. The method of any one of claims 1 to 32, comprising administering a second compound that increases satiety, reduces food intake, loses or maintains weight, or performs one of these combinations. It further comprises a method. 36. The method of claim 35, wherein the second compound is an exendin or an agonist thereof, amylin or an agonist thereof, PYY or an agonist analog thereof, leptin, oxyntomodulin, or At least one compound selected from the group consisting of cholecystokinin (CCK). Use of an endothelin or endothelin agonist in an amount effective to reduce food intake to produce a drug for a patient who desires or needs to reduce food intake. Use of an endothelin or endothelin agonist in an amount effective for weight loss to prepare a weight loss drug for a patient who desires or needs weight loss. Use of an endotherin or endothelin agonist in an amount effective to treat obesity, for the manufacture of an anti-obesity drug for a patient who desires or needs treatment of obesity. Use of an endotherin or endoterin agonist at a dose effective to treat a metabolic disorder, for the manufacture of a medicament for treating or preventing metabolic disorders for a patient in need or need of treating or preventing a metabolic disorder. 41. The use of claim 40, wherein the metabolic disease is obesity, diabetes mellitus, insulin-resistant syndrome, syndrome-X, or other hypertrophic disease. 42. The method of any one of claims 37-41, wherein the patient has a future weight gain associated with a disease or condition such as obesity, overweight, a patient in need of weight loss or weight gain caused by drugs. Use, characterized in that the patient in need of suppression. 42. The use according to any one of claims 37 to 41, wherein said endoterin is at least one of endoterin-1, endoterin-2, or endoterin-3. 44. The use of claim 43, wherein said endoterin is a sequence having at least 75% sequence homology with endoterin-1 or SEQ ID NO: 1. 44. The use of claim 43, wherein said endoterin is a sequence having at least 75% sequence homology with endoterin-2 or SEQ ID NO: 7. 44. The use according to claim 43, wherein said endoterin is a sequence having at least 75% sequence homology with endoterin-3 or SEQ ID NO. 42. The method according to any one of claims 37 to 41, wherein the endothelin agonist is an analog of SEQ ID NO: 1, SEQ ID NO: 7, or SEQ ID NO: 10 that does not include substitutions, deletions or additions of at least 5 amino acids. Use. 48. The use of claim 47, wherein the analog does not comprise a substitution of at least 5 amino acids. 42. The use according to any one of claims 37 to 41, wherein said endoterin is endoterin-1 selected from at least one of the groups consisting of SEQ ID NOs: 1-6. 42. The use according to any one of claims 37 to 41, wherein said endoterin is endoterin-2 selected from at least one of the group consisting of SEQ ID NOs: 7-9. 42. The use according to any one of claims 37 to 41, wherein said endoterin is endoterin-3 selected from at least one of the group consisting of SEQ ID NOs: 10-13. 42. The use of any one of claims 37 to 41, wherein the endothelin agonist is an endothelin agonist that contains no more than 35 amino acids. The use of any one of claims 43-52, wherein the endothelin agonist comprises a C terminus having the amino acid sequence FCHLDIIW. The use of any one of claims 43-52, wherein the endothelin agonist comprises a C terminus having the amino acid sequence FAHLDIIW. 43. The method according to any one of claims 37 to 41, wherein the endothelin agonist is N-cis-2,6-dimethylpiperidinocarbonyl-L-γ-methylleucine-D-1-methoxycarbonyltrip Topanyl-D-norleucine. 42. The use of any one of claims 37 to 41, wherein the endothelin agonist is [Ala ^{1, 3, 11, 15} ] endothelin-1 (SEQ ID NO: 14). 42. The method of any one of claims 37-41, wherein the endothelin agonist is a fragment of endothelin-1, endothelin-2, endothelin-3, wherein the fragment binds to and activates an ET _A or ET _B receptor. Use, characterized in that. 59. The use of claim 57, wherein the fragment does not comprise at least seven amino acid deletions. 42. The use of any one of claims 37-41, wherein said endothelin agonist does not comprise at least 10 amino acid substitutions, additions or deletions compared to any one of SEQ ID NOs: 1, 7 and 10. 59. The use of claim 57 or 58, wherein the endothelin agonist is (N-succinyl- [Glu ⁹ , Ala ^11,15 ] ^-endoterin -1 fragment 8-21. 59. The use of claim 57 or 58, wherein the endothelin agonist is (N-acetyl- [Ala ^11,15 ] ^-endoterin -1 fragment 6-21. 42. The use of any one of claims 37 to 41 wherein the endothelin agonist is sarafotoxin. 63. The use of claim 62, wherein the sarapotoxin has an amino acid sequence selected from the group consisting of SEQ ID NOs: 15, 16, 17, 18, 31 and 32. 63. The use of claim 62, wherein the sarapotoxin has an amino acid sequence having at least 75% sequence homology with SEQ ID NOs: 15, 16, 17, 18, 31, and 32. 65. The use of any one of claims 62 to 64, wherein the sarapotoxin comprises a C terminus having the amino acid sequence FCHQDVIW. 42. The use of any one of claims 37 to 41, wherein the endothelin agonist is demseptin or adenoregulin. 67. The use of claim 66, wherein the dumbceptin or adenoregulin has an amino acid sequence selected from the group comprising SEQ ID NOs: 20-30 and SEQ ID NOs: 33-64. 67. The use of claim 66, wherein the dumbceptin or adenoregulin has an amino acid sequence having at least 75% sequence homology to any one of SEQ ID NOs: 20-30 and SEQ ID NOs: 33-64. 69. The use of any one of claims 37-68, wherein the endothelin or endothelin agonist is an ET _B receptor agonist but not an ET _A receptor agonist. 70. The use of any one of claims 37-68, wherein the endothelin or endothelin agonist binds to and activates the ET _B receptor as compared to the ET _A receptor. 39. The use of any one of claims 37-38 for administering a second compound that increases satiety, reduces food intake, reduces or maintains weight, or performs one of these combinations. For further use. 72. The method of claim 71, wherein the second compound is an exendin or an agonist thereof, amylin or an agonist thereof, PYY or an agonist analog thereof, leptin, oxyntomodulin, or And at least one compound selected from the group consisting of cholecystokinin (CCK). 73. The patient of any one of claims 1 to 72 wherein the patient is obese, overweight, a patient in need of weight loss, a patient in need of weight maintenance, or in need of inhibiting future weight gain. Use or method characterized in that the patient. 74. The use or method of Claim 73, wherein the gain in body weight is caused by the drug.

Images